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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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How specific is CRISPR/Cas9 really?

Henriette O'Geen1, Abigail S Yu1, David J Segal1

  • 1Genome Center, Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616, USA.

Current Opinion in Chemical Biology
|October 31, 2015
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 gene editing shows conflicting reports on its specificity. This review deciphers claims about its precision for research and therapeutic applications.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • RNA-guided nucleases, including CRISPR/Cas9, are vital tools in research and therapy.
  • Early studies produced conflicting data regarding the precision of CRISPR/Cas9 genome editing.
  • This led to the development of strategies to enhance specificity and detect off-target mutations.

Purpose of the Study:

  • To review and interpret the contradictory findings on CRISPR/Cas9 system specificity.
  • To clarify the precision of CRISPR/Cas9 for genome engineering applications.

Main Methods:

  • Review of existing scientific literature on CRISPR/Cas9 specificity.
  • Analysis of studies reporting on both off-target events and high specificity.
  • Interpretation of conflicting data to resolve discrepancies.

Main Results:

  • Early concerns about CRISPR/Cas9 promiscuity contrasted with later findings of high specificity.
  • Various methods have been developed to assess and improve editing precision.
  • The field has grappled with understanding the nuances of CRISPR/Cas9 targeting.

Conclusions:

  • CRISPR/Cas9 can be a highly specific genome-editing tool when properly utilized.
  • Understanding and mitigating off-target effects remain critical for safe therapeutic development.
  • Further research is needed to fully elucidate the factors influencing CRISPR/Cas9 specificity.